Understanding and manipulating chronic Helicobacter pylori to enhance treatment

我们建议的研究重点是确定限制慢性病原体HELI对抗生素敏感性的因素。 幽门螺杆菌。有证据表明，慢性幽门螺杆菌很难用抗生素治愈，因为它处于一种 缓慢的生长状态至少部分受胃酸控制。幽门螺杆菌的治疗依赖于通过In- 包括被称为质子泵抑制剂(PPI)的强抗酸剂。PPI阻止了酸液的生产，提高了气孔- ACh pH，促进幽门螺杆菌生长。细菌的生长可以让标准的抗生素发挥更好的作用。有一个 在我们对幽门螺杆菌慢性生长状态的确切性质的理解上存在差距，例如它的代谢有多活跃- 问题是，酸是否是唯一的生长抑制物，以及幽门螺杆菌在什么代谢条件下生长 PPI治疗。这一信息很重要，因为幽门螺杆菌感染是在慢性状态下治疗的。MIL- 全世界和美国有成群结队的人感染幽门螺杆菌，并患有与之相关的疾病- 溃疡和胃癌。胃癌是全球第四大癌症死亡原因。幽门螺杆菌是一种 这是一个持续的问题，因为发达国家的发病率已经稳定下来。此外，目前的治疗方法 治愈幽门螺杆菌感染失败的频率令人无法接受：美国最近的估计发现 目前的治疗方案无法治愈20-25%的感染者。本AP的总体目标是- 复制是为了了解幽门螺杆菌的慢性生长状态，并使用这些信息来设计 促进生长，从而提高抗生素敏感性。我们的中心假设，基于已发表的和初步的 数据显示，大多数慢性状态的幽门螺杆菌处于极缓慢的生长模式，受到 酸、翻译缺乏症和营养限制的组合。在目标1中，我们将使用H。 幽门螺杆菌突变体和小鼠模型填补了我们对幽门螺杆菌慢性生长状态和 生长速度，这些参数如何受PPI影响，以及PPI后倍增是否需要乳酸 像早期乘法一样的利用率。此外，我们测试是否增加关键的碳源，如 乳酸可促进幽门螺杆菌的慢性生长和抗生素治疗。在目标2中，我们以初步数据为基础 表现出生长缓慢的幽门螺杆菌表现出显著的翻译抑制，包括通过增加核糖体- MAL沉默因子RsfS。我们使用分子生物学和生物化学来填补我们对Rsfs理解上的空白。 功能，并表征如何控制RsfS的表达，以及其他翻译抑制。 TORS，控制翻译，影响长期殖民。这项拟议的研究在假设方面是创新的。 除了酸，幽门螺杆菌的慢性缓慢生长还受到信号的促进，而且知道和靶向 这些将促进更好的治疗方法。这项拟议的研究具有重要意义，因为它将为 控制慢性生长的方法，并提供了提高幽门螺杆菌对抗生素敏感性的新方法。这个 这项研究产生的长期结果将提供洞察力，为改进奠定基础 将这些微生物推向抗生素敏感状态的治疗方法。

Our proposed research focuses on defining factors that limit antibiotic sensitivity of the chronic pathogen Heli- cobacter pylori. Evidence suggests that chronic H. pylori is difficult to cure with antibiotics because it is in a slow growth state controlled at least in part by stomach acid. H. pylori treatments rely on removing acid by in- cluding strong antacids called proton pump inhibitors (PPI). The PPI blocks acid production, raises the stom- ach pH, and promotes H. pylori growth. Bacterial growth allows standard antibiotics to work better. There is a gap in our understanding of the exact nature of the H. pylori chronic growth state, e.g. how active its metabo- lism is, whether acid is the only growth inhibitor, and what type of metabolism H. pylori deploys to grow after PPI treatment. This information is important because H. pylori infections are treated at the chronic state. Mil- lions of people worldwide and in the U.S. are infected by H. pylori and suffer from its associated diseases— ulcers and gastric cancer. Gastric cancer is the fourth leading cause of cancer deaths worldwide. H. pylori is an on-going problem, as the incidence has stabilized in the developed world. Furthermore, current therapies to cure H. pylori infection fail with unacceptable frequency: recent estimates in the United States have found that 20-25% of infected individuals are not cured by the current therapeutic regime. The overall objective of this ap- plication is to understand the H. pylori chronic growth state and use this information to design approaches that enhance growth and therefore antibiotic sensitivity. Our central hypothesis, based on published and preliminary data, is that the majority of chronic-state H. pylori are in an extreme slow growth mode, limited by a combination of acid, translational deficiency, and nutrient restriction. In Aim 1, we will use a combination of H. pylori mutants and mouse models to fill gaps in our understanding of the H. pylori chronic growth state and growth rate, how these parameters are affected by PPI, and whether post-PPI multiplication requires lactate utilization as early stage multiplication does. Additionally, we test whether increasing key carbon sources like lactate enhances H. pylori chronic state growth and antibiotic cure. In Aim 2, we build on preliminary data showing slow growth H. pylori display significant translational repression, including by increase in the riboso- mal silencing factor RsfS. We use molecular biology and biochemistry to fill gaps in our understanding of RsfS function in general, and to characterize how controlled RsfS expression, as well as other translational inhibi- tors, controls translation and affect chronic colonization. The proposed research is innovative in its hypothesis that H. pylori chronic slow growth is promoted by signals in addition to acid, and that knowing and targeting these will promote better cures. The proposed research is significant because it will provide new insights into ways that chronic growth is controlled and provide new ways to enhances H. pylori antibiotic sensitivity. The long-term outcomes generated by this research will provide insights that will lay the groundwork for improved therapies that push these microbes into an antibiotic-sensitive state.